It is reported that up to 40% of neonates undergoing cardiac surgery suffer a major thrombosis event. Pediatric patients who are at increased risk for thrombosis due to either congenital or acquired hypercoagulability require individualized anticoagulation management. Presence of a thrombophilia influences clinical decision making regarding timing and duration of anticoagulation, particularly in children with venous thrombosis. The duration of therapy is guided by the number of risk factors and thrombophilic conditions. Hypercoagulability panel screening allows risk stratification for therapeutic management. Screening for hypercoagulability is indicated in pediatric patients with a family history of thrombosis, or in those who have suffered a major thrombotic or thromboembolic event. However, a large number of patients develop thrombosis without meeting current criteria for screening. Currently available screening tests are expensive, time-consuming and ordered individually, requiring large volumes of blood sample. These issues make routine screening for a full hypercoagulability panel clinically impractical, especially in pediatric patients. Hence, thre is a compelling need for an inexpensive hypercoagulability laboratory panel that requires very small volumes of blood, has a rapid turn-around time and can be applied to a larger number of potentially at-risk patients. Such a diagnostic panel is possible by employing a digital microfluidic system. Hypercoagulability tests on a digital microfluidic cartridge will enable portable automation, rapid turn-around time, and multiplexing several assays, including different types of assays (immunoassays and functional) utilizing about 100 nL sample per test. A disposable cartridge for hypercoagulability panel testing using digital microfluidic manipulation of droplets will be developed. The ultimate goal is the development of a single platform that can perform multiplexed immunoassays or enzyme-linked immunosorbent assay (ELISA) and functional assays using a finger prick whole blood sample to determine hypercoagulability risk, with initial emphasis on pediatric patients undergoing such testing. In Phase I, we propose to integrate the multiplexed immunoassays (Anti-thrombin III, Protein C and Plasminogen Activator Inhibitor-1) with functional assays (Anti-thrombin III, Protein C, Plasminogen, FVIII and Thrombin generation) allowing for a complete thrombophilia panel on a single disposable cartridge. In Phase II, on-cartridge reagent storage, comprehensive quality control, and other necessary activities for commercialization of a product will be developed.